H.C. Chauhan

Comparison of the standard AGID test and competitive ELISA for detecting bluetongue virus antibodies in camels in Gujarat, India.

The performance of the standard agar gel immunodiffusion (AGID) test and the competitive enzyme-linked immunosorbent assay (cELISA) for the detection of serum antibody against bluetongue virus (BTV) in clinically healthy and diseased camels in Gujarat state were compared. Out of 176 sera tested, 22 (12.5%) and 34 (19.3%) were positive for group-specific bluetongue antibodies by AGID and cELISA, respectively. Maximum seropositivities of 18.0% by AGID and 25.8% by cELISA were recorded in the Kutchhi breed, and of 6.9% and 12.6%, respectively, in the Marwari breed. The seroprevalence detected by AGID and cELISA in clinically healthy and diseased camels did not differ significantly with regard to bluetongue disease in these breeds.

Comparison of molecular and microscopic technique for detection of Theileria annulata from the field cases of cattle

Aim: Tropical theileriosis is fatal hemoprotozoal disease of dairy animals caused by Theileria annulata. The aim of the present study was to detect the T. annulata and comparison of results of molecular and microscopic techniques. Materials and Methods: A total of 52 blood samples were collected from the cattle suspected for theileriosis across the Banaskantha district. All the samples were screened for theileriosis using Giemsa’s staining technique and polymerase chain reaction (PCR). Results: Total of 17 (32.69%) and 24 (46.15%) samples were found positive for theileriosis by microscopic examination and PCR test, respectively. It revealed that the study area is endemic for theileriosis, and the microscopic technique has 70.83% sensitivity and 100% specificity with respect to PCR technique. Conclusion: It may be concluded from the present study that the PCR is comparatively sensitive technique than microscopic examination and may be recommended to use in the field for screening of theileriosis in the study area, where a high prevalence of diseases have been reported due to intensive dairy farming.

Genotyping of Indian antigenic, vaccine, and field Brucella spp. using multilocus sequence typing

INTRODUCTION Brucellosis is one of the most important zoonotic diseases that affects multiple livestock species and causes great economic losses. The highly conserved genomes of Brucella, with > 90% homology among species, makes it important to study the genetic diversity circulating in the country. METHODOLOGY A total of 26 Brucella spp. (4 reference strains and 22 field isolates) and 1 B. melitensis draft genome sequence from India (B. melitensis Bm IND1) were included for sequence typing. The field isolates were identified by biochemical tests and confirmed by both conventional and quantitative polymerase chain reaction (qPCR) targeting bcsp 31Brucella genus-specific marker. Brucella speciation and biotyping was done by Bruce ladder, probe qPCR, and AMOS PCRs, respectively, and genotyping was done by multilocus sequence typing (MLST). RESULTS The MLST typing of 27 Brucella spp. revealed five distinct sequence types (STs); the B. abortus S99 reference strain and 21 B. abortus field isolates belonged to ST1. On the other hand, the vaccine strain B. abortus S19 was genotyped as ST5. Similarly, B. melitensis 16M reference strain and one B. melitensis field isolate were grouped into ST7. Another B. melitensis field isolate belonged to ST8 (draft genome sequence from India), and only B. suis 1330 reference strain was found to be ST14. CONCLUSION The sequences revealed genetic similarity of the Indian strains to the global reference and field strains. The study highlights the usefulness of MLST for typing of field isolates and validation of reference strains used for diagnosis and vaccination against brucellosis.

Sero-Epidemiology of Camel (Camelus dromedarius) brucellosis

Brucellosis is an emerging zoonotic bacterial disease of ruminants and also reported in camels. Camels are one of the most important sources of livelihood for the poor nomadic population in Gujarat. The present study was aimed to determine the brucella specific antibodies in camel using RBPT and i-ELISA. On screening of 658 serum samples, 131(19.90%) and 78 (11.85%) samples found to be positive by RBPT and i-ELISA, respectively. Prevalence rate of brucellosis in different categories viz. herd size, physiological status, sex, district, region and breed were calculated. Susceptibility of brucellosis in different categories was also compared by using chi-square test.

Phenotypic and Genotypic Identification of Methicillin-Resistant Coagulase Negative Staphylococcus spp. Isolated from Bovine Mastitis

India ranks first in the world in milk production and dairying in India are a classic example of production by masses rather than mass production. Among the different food sectors, the growth in dairy sector has been commendable. The rate of growth in milk production in India is also substantially higher (3.60 %) than the world average of 1.50 per cent. However, the total projected demand of milk by the year 2030 would be about 200 million tonnes, depending on assumptions about income, population, urban growth, and expenditure elasticity parameters, which would imply an annual increase of around 4 million tonnes during the next two decades (N.D.R.I., 2011). Among the several barriers in achieving the production targets, mastitis continues to remain as a challenging impediment, since the affected quarters may have 30.00 per cent less productivity and dairy animals may lose about 15.00 per cent production (Radostitis et al., 2007). In India, the overall economic loss due to mastitis is estimated to be Rs. 7165.51 crores (Bansal and Gupta, 2009). Despite intense research and control programmes, bovine mastitis has remained a major economic problem of the dairy industry (Denis et al., 2009). International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 7 Number 03 (2018) Journal homepage: http://www.ijcmas.com

Metagenomic of clinically diseased and healthy broiler affected with respiratory disease complex

In recent past, the respiratory infection has emerged as a great challenge to the poultry farmers. Various pathogens including Avian pneumovirus (APV), Avian influenza virus (AIV), Infectious bronchitis virus (IBV) and Newcastle disease virus (NDV), Avibacterium paragallinarum, Ornithobacterium rhinotracheale (ORT), Mycoplasma synoviae (MS), Mycoplasma gallisepticum (MG) and Avian pathogenic Escherichia coli (APEC) are involved in the respiratory disease complex in birds [1], [2] (Bradbury, 1984; Roussan et al., 2008). Hence, respiratory disease complex is the most serious disease affecting to poultry and causes heavy economic losses in the poultry industry worldwide [3] (Murthy et al., 2008). In recent years, metagenomics is powerful analyzing tool for detection of pathogens directly from clinical samples without any prior knowledge of the organism in a given sample [4], [5] (Schuster, 2008; Pereira et al., 2010). High throughput Next-Generation-Sequencing technology was used for sequencing the isolated genomic DNA. These data provides an insight about taxonomic and functional status of microorganisms responsible for causing respiratory infection in broiler. The data of these metagenome are available in the BioSample Submission Portal as Bioproject PRJNA339659 and SRA accession number SRR5997823, SRR5992854, SRR6037376, SRR6024702, SRR6012248 and SRR6008913.

Detection of Bluetongue Virus Antigen from Livestock of Gujarat State

Bluetongue (BT) is an infectious, non-contagious disease of domestic and wild ruminants. Bluetongue virus (BTV) causes severe disease in sheep, which is transmitted by insect vector belonging to Culicoides spp. It is particularly a viral disease of sheep, occasionally affecting cattle, buffaloes, goats, camels and other wild ruminants. Out of 377 (364-blood, 5-spleen and 8-pooled Culicoides) samples 110 (29.18%) and 28 (7.42%) were found positive for BTV antigen by s-ELISA and BT-AGID respectively. Specieswise incidence by s-ELISA recorded was 48.20 per cent in sheep, 57.14 per cent in goats and 2.60 per cent in cattle however, none of the blood sample found positive from buffalo and camel. Specieswise incidence by BT-AGID recorded was 12.23 per cent in sheep and 15.71 per cent in goats however, none of the blood sample found positive for BTV antigen from cattle, buffalo and camel. Higher incidence seen in goats by both the test. s-ELISA proved to be the most sensitive in detecting BTV antigen than BT-AGID. Considering s-ELISA as the reference test, the relative sensitivity, specificity and overall agreement between both the tests were 25.45 per cent, 100 per cent and 78.24 per cent respectively.

Ideal Sample for Isolation of Pasteurella multocida and Susceptibility of Field Isolates Against Commonly Used Antibiotics

Haemorrhagic septicaemia (HS) is an acute pasteurellosis manifested by an acute and highly fatal septicaemia may cause high mortality. Therefore, early diagnosis and effective treatment may help to control the mortality. The present study was aimed to evaluate the suitability of various samples (blood, bone marrow, liver and lung) for isolation of Pasteurella multocida and their susceptibility against various antibiotics. Total 52 samples [blood (n= 25), bone marrow (n=9), liver (n=9) and pneumonic lung (n=9)] were collected from HS-suspected animals and subjected for bacterial isolation after confirming the disease using microscopy. Total 10 isolates [2 (blood), 2 (bone marrow), 1 (liver) and 5 (lung)] were isolated and confirmed by morphological characteristics, biochemical test and Polymerase Chain reaction (PCR). Routinely used antibiotics were screened for their sensitivity against these isolates. Based on the present study, it may be concluded that the lung tissue sample may be the ideal sample for P. multocida isolation, whereas, gentamicin and chloramphenicol are effective antibiotics against all the isolates.

Seroprevalence of bluetongue in dromedaries

Bluetongue (BT) is an infectious, non-contagious, arthropod-borne viral disease, mainly of sheep but many domestic and wild animals are also affected by this disease. A serological study was aimed at the detection of BTV antibodies by Agar Gel Immunodiffusion test (BT-AGID) and competitive Enzyme Linked Immunosorbent Assay (c-ELISA) in dromedaries of North Gujarat and Kachchh regions. Out of 533 serum samples, the BT-AGID test detected antibodies against BTV in 83 cases (15.57%) while c-ELISA test was positive in 136 cases (25.51%).

Isolation, Identification and Antibiogram of Staphylococcus aureus Isolated from Bovine Mastitis

In the present study, out of 255 milk samples (185 from subclinical and 70 from clinical cases) screened for Staphylococcus aureus, 53 isolates were obtained giving an overall incidence of 20.78%. The incidence of S. aureus from subclinical mastitis was 8.11% (15/185) and from clinical mastitis was 54.29% (38/70). Species-wise incidence was 30.77% (16/52) in buffaloes and 18.23% (37/203) in cows. All 53 isolates were identified on the basis of colony characters on nutrient agar, staining and biochemical characters namely catalase production, oxidise test, maltose fermentation test and ohosphatase test. Out of 53 isolates, 49 (92.45%) isolates showed coagulase production in tube coagulase test. Number of isolates showing alpha, beta, gamma and alpha-beta haemolysin production were 18 (33.96%), 26 (49.06%), 4 (7.55%) and 5 (9.43%) on sheep blood agar, respectively. All the 53 S. aureus isolates were confirmed genotypically by 23S rRNA ribotyping in which a species-specific amplicon of 1, 250 bp was obtained. Antibiotic sensitivity patterns of all these 53 S. aureus isolates revealed that the isolates were most sensitive to gentamicin (94.34%), whereas penicillin (94.34%) and ampicillin (86.79%) were highly resistant to S. aureus.